Tuning the conformational properties of the prion peptide
Chai-Chi Ho†$, Lily Y.-L. Lee†$, Kuo-Ting Huang‡, Chun-Cheng Lin‡#, Mei-yun Ku†,
Chien-Chih Yang§, Sunney I. Chan‡, Ruei-Lin Hsu† and Rita P.-Y. Chen†*
†
Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan, R. O. C
‡
#
Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan, R. O. C
Department of Chemistry, National Tsing Hua University, Hsinchu 30013, Taiwan,
R.O.C.
§
Department of Biochemical Science and Technology, National Taiwan University,
Taipei 106, Taiwan, R. O. C
Supporting Information
Table of Content
Table S1. Amino acid sequences of the synthesized peptides.
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Figure S1. Comparison of 2D-TOCSY spectra of PrP(108-144), K110G, and R136G
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Figure S2. Comparison of 1D-spectra of PrP(108-144), R136G, and K110G
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Figure S3. TEM images of the PrP(108-144) fibrils and the fibrils formed from
S135--GlcNAc seeded with the PrP(108-144) fibrils.
Supporting information - 1
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Table S1. Amino acid sequence of the synthesized peptides. The difference from
the wildtype peptide is highlighted in red.
Name
Sequence
PrP(108-144)
Ac-NMKHMAGAAAAGAVVGGLGGYMLGSAMSRPMMHFGND-NH2
S135--GalNAc
Ac-NMKHMAGAAAAGAVVGGLGGYMLGSAMS*RPMMHFGND-NH2
S135--GlcNAc
Ac-NMKHMAGAAAAGAVVGGLGGYMLGSAMS*RPMMHFGND-NH2
S135--GalNAc
Ac-NMKHMAGAAAAGAVVGGLGGYMLGSAMS*RPMMHFGND-NH2
S135--GlcNAc
Ac-NMKHMAGAAAAGAVVGGLGGYMLGSAMS*RPMMHFGND-NH2
R136G
Ac-NMKHMAGAAAAGAVVGGLGGYMLGSAMSGPMMHFGND-NH2
K110G
Ac-NMGHMAGAAAAGAVVGGLGGYMLGSAMSRPMMHFGND-NH2
R136/ S135--GalNAc
Ac-NMKHMAGAAAAGAVVGGLGGYMLGSAMS*GPMMHFGND-NH2
* glycosylated site
Supporting information - 2
Figure S1. Comparison of 2D-TOCSY of PrP(108-144), K110G, and R136G.
Peptides were dissolved in 20 mM H3PO4 and 10 % D2O to a final concentration of
1mM. A 1/100 volume of sodium 3-(trimethylsilyl)-propionic-2, 2, 3, 3-d4 acid
solution (0.75 % in D2O) was added as an internal reference. An 80 millisecond
mixing time was used to record 2D-TOCSY on a Bruker AVANCE 600 AV NMR
spectrometer at 298K. The H versus H crosspeaks of K110 do not show in the
spectrum of K110G due to K110→G mutation. The intra-residue crosspeaks of
M109 and H111 are also affected by the K110G mutation. The H versus H
crosspeaks of R136 do not show in the spectrum of R136G due to R136→G mutation.
R136G is prone to associate and hence the crosspeak intensity of its spectrum is
weaker than those of the other two peptides.
Supporting information - 3
Figure S2. Comparison of 1D-spectra of PrP(108-144), K110G, and R136G.
Peptides were dissolved in 20 mM H3PO4 and 10 % D2O to a final concentration of
1mM. A 1/100 volume of sodium 3-(trimethylsilyl)-propionic-2, 2, 3, 3-d4 acid
solution (0.75 % in D2O) was added as an internal reference. The spectra were
recorded on a Bruker AVANCE 600 AV NMR spectrometer at 298K.
Supporting information - 4
Figure S3. TEM images of the PrP(108-144) fibrils and the fibrils formed from
S135--GlcNAc seeded with the PrP(108-144) fibrils. The images were recorded in
a Tecnai G2 Spirit TWIN electron microscope (FEI Company). The corresponding
length of scale bar is shown in each figure.
Supporting information - 5